Under normal operation conditions, granular activated carbon (GAC) employed in drinking water treatment plants (DWTPs) for natural organic matter (NOM) removal can be colonised by microorganisms which can eventually establish active biofilms. The formation of such biofilms can contribute to NOM removal by biodegradation, but also in clogging phenomena that can make necessary more frequent backwashes. Biofilm occurrence and evolution under full-scale-like conditions (i.e. including periodic backwashing) are still uncertain, and GAC filtration is usually operated with a strong empirical component. The aim of the present study was to assess the formation and growth, if any, of biofilm in a periodically backwashed GAC filter. For this purpose, an on-site pilot plant was assembled and operated to closely mimic the GAC filters installed in the DWTP in Sant Joan Despí (Barcelona, Spain). The study comprised a monitoring of both water and GAC cores withdrawn at various depths and times throughout 1 year operation. The biomass parameters assessed were total cell count by confocal laser scanning microscopy (CLSM), DNA and adenosine triphosphate (ATP). Visual examination of GAC particles was also conducted by high-resolution field emission scanning electron microscopy (FESEM). Additionally, water quality and GAC surface properties were monitored. Results provided insight into the extent and spatial distribution of biofilm within the GAC bed. To sum up, it was found that backwashing could physically detach bacteria from the biofilm, which could however build back up to its pre-backwashing concentration before next backwashing cycle.